Biolegend Cell Surface Immunofluorescence Staining Protocol uses 15 ml of cell staing buffer to resuspend the cell. Is it necessary? Here is the link: https://www.biolegend.com/protocols/cell-surface-immunofluorescence-staining-protocol/4283/
It is a washing step allowing to obtain clean cells for future procedures. You get rid of extracellular proteins, medium components etc by first diluting them in excess volume, and then spinning the sample to keep the pellet and discard the supernatant.
May be it is total volume for all washing steps (but not that big volume for one washing). For re-suspension the cell, the volume should be much less than 15 ml; for me it is only from 0.5-1 ml depending on cell density.
Dear Dr. Di Yang ,
Don't bother about 15 ml. Based on the amount of cell suspension you have, add 5 times more PBS + 2% FBS and wash . It works beautifully. I have done >1000 times. Feel free to knock me for further assistance @ [email protected]
BR,
Babu M.
Normally to safe the chemical, we use the glass tube (4 ml) with cover slip for cell staining washing. You only add 0.5- 1 ml each for cell pellet, gently voltex, then centrifuge; 3 times washing. Then you add FACS buffer for measurement.
Hope this information could help you.
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